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Live-Cell Imaging of Microglia in Organotypic Brain Slices Using Microcontact Printing
Björn Y P Richardsen1, Christian Humpel1
1Laboratory of Psychiatry and Experimental Alzheimer's Research, Department of Psychiatry and Psychotherapy, Medical University of Innsbruck, Anichstr. 35, A-6020 Innsbruck, Austria.
Abstract:
Microglia are brain immune cells that phagocytose cell debris and beta-amyloid plaques in patients with Alzheimer's disease. They develop from round amoeboid cells into ramified microglia or large macrophages, which can be studied in three-dimensional organotypic mouse brain slices. In a recent publication, we showed for the first time that we can track GFAP+ astrocytes and laminin+ vessels in organotypic brain slices using live-cell imaging . The aim of the present study was to use microcontact printing on organotypic brain slices to label microglia with Iba1 and CD11b antibodies and visualise them through live-cell imaging. We show that microglia can be easily labelled with antibodies and tracked via live-cell fluorescence microscopy for up to 20 days. Incubation in lipopolysaccharide (LPS) or granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulates the migration of round amoeboid microglia, whereas interleukin-10 induces their differentiation into ramified forms. Taken together, we show the first-time live cell imaging of microglia in organotypic mouse brain slices using microcontact printing.
Insights
Researchers visualize microglia, the brain's immune cells, in mouse brain slices for up to 20 days. This live-cell imaging technique tracks microglia's response to stimuli, aiding Alzheimer's disease research.
Area of Science:
- Neuroscience
- Immunology
- Cell Biology
Background:
- Microglia are crucial brain immune cells involved in clearing debris and beta-amyloid plaques, particularly relevant in Alzheimer's disease.
- Microglia exhibit dynamic morphological changes, transitioning from amoeboid to ramified forms or macrophages, which can be studied in organotypic brain slice models.
- Previous work established live-cell imaging of astrocytes and vessels in these slices, setting the stage for microglia visualization.
Purpose of the Study:
- To develop and validate a method for live-cell imaging of microglia in organotypic mouse brain slices.
- To utilize microcontact printing for antibody labeling of microglia (Iba1 and CD11b) for enhanced visualization.
- To observe microglia's dynamic behavior and responses to specific stimuli in a controlled ex vivo environment.
Main Methods:
- Organotypic mouse brain slices were prepared for microscopy.
- Microcontact printing was employed to apply Iba1 and CD11b antibodies for microglia labeling.
- Live-cell fluorescence microscopy was used to track labeled microglia over time (up to 20 days).
- Microglia responses to lipopolysaccharide (LPS), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-10 (IL-10) were assessed.
Main Results:
- Microglia were successfully labeled with antibodies and visualized using live-cell fluorescence microscopy for an extended period of 20 days.
- Stimulation with LPS or GM-CSF induced migration of round amoeboid microglia.
- Interleukin-10 treatment promoted the differentiation of microglia into ramified forms.
- This study presents the first instance of live-cell imaging for microglia in organotypic brain slices using microcontact printing.
Conclusions:
- A robust method for live-cell imaging of microglia in organotypic brain slices has been established.
- Microcontact printing facilitates efficient antibody labeling and tracking of microglia.
- The technique allows for the observation of microglia's dynamic morphological and migratory responses to various stimuli, providing insights into their role in neurological conditions like Alzheimer's disease.
